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Acta Agron Sin ›› 2017, Vol. 43 ›› Issue (09): 1410-1414.doi: 10.3724/SP.J.1006.2017.01410

• RESEARCH NOTES • Previous Articles    

Differential Expression and Assembly Mode of Glutamine Synthetase Isoen-zymes in Different Tissues and Organs of Maize

WANG Xiao-Chun1,2,3,ZHANG Hao-Ran3,WEI Yi-Hao1,JIA Xi-Ting3,GU Ming-Xin3,MA Xin-Ming1,*   

  1. 1 Collaborative Innovation Center of Henan Grain Crops / Henan AgriculturalUniversity, Zhengzhou 450002, China; 2 State Key Laboratory of Wheat and Maize Crop Science in China / Henan AgriculturalUniversity, Zhengzhou 450002, China; 3 Department of Biochemistry, College of Life Science / Henan AgriculturalUniversity, Zhengzhou 450002, China
  • Received:2016-12-21 Revised:2017-04-20 Online:2017-09-12 Published:2017-05-08
  • Contact: Ma Xinming, E-mail: xinmingma@126.com, Tel:13937100780 E-mail:xiaochun.w@163.com
  • Supported by:

    The study was supported by the National Key Research and Development Program of China (2016YFD0300205) and State Key Laboratory of Wheat and Maize Crop Science (39990047).

Abstract:

Glutamine synthetase (GS) is a key enzyme in nitrogen assimilation and recycling in cereals. In this study, the expression characteristics of GS isoenzymes in different tissues and organs of maize in grain-filling period were analyzed, and the assembly of GS isoenzymes were indentified. The GS isoforms expressed differentially in different organs were shown by Western-blot obviously; GS1 with a molecular weight of about 40 kD expressed highly in all tissues, and GS1 with a molecular weight of about 39 kD was merely expressed in the node of ear position and pedical, and GS2 with a molecular weight of about 44 kD was weakly expressed in the photosynthtic tissue such as leaf. With a modified blue naive PAGE (BNE) technique and in-gel activity analysis, the size of GS holoenzyme was calibrated; combined the 2-D gel with western-blot analysis, the subunits composition of GS isoenzymes were identified. Three GS isoenzymes with different sizes were identified in maize. GS2 holoenzyme was about 460 kD and likely a decamer, GS1 holoenzyme existed two kinds of assembly state, one was about 410 kD and likely a decamer, another was about 240 kD and more likely a pentamer; therefore, the expression of GS isoenzymes exists diversity in maize.

Key words: Maize, Glutamine synthetase (GS), Expression, Blue native PAGE (BNE), Assembly

[1] Bernard S M, Habash D Z. The importance of cytosolic glutamine synthetase in nitrogen assimilation and recycling. New Phytol, 2009, 182:608–620
[2] McNally SF, Hirel B, Gadal P, Mann AF, Stewart GR. Glutamine synthetases of higher plants: evidence for a specific isoform content related to their possible physiological role and their compartmentation within the leaf. Plant Physiol, 1983, 72: 22–25
[3] Hirel B, Gadal P. Glutamine synthetase isoforms in pea leaves: intracellular localization. Zeitschrift Fü Pflanzenphysiologie, 1981, 102: 315–319
[4] Hirel B, Lea P J. Ammonia Assimilation. Plant Nitrogen. Springer Berlin Heidelberg, 2001. pp 79–99
[5] Zozaya-Hinchliffe M, Potenza C, Ortega J L, Sengupta-Gopalan C. Nitrogen and metabolic regulation of the expression of plastidic glutamine synthetase in alfalfa (Medicago sativa). Plant Sci, 2005, 168: 1041–1052
[6] Cren M, Hirel B. Glutamine synthetase in higher plants regulation of gene and protein expression from the organ to the cell. Plant Cell Physiol, 1999, 40: 1187–1193
[7] Robert F M, Wong P P. Isozymes of glutamine synthetase in Phaseolus vulgaris L. and Phaseolus lunatus L. root nodules. Plant Physiol, 1986, 81: 142–148
[8] Carvalho H, Pereira S, Sunkel C, Salema R. Detection of a cytosolic glutamine synthetase in leaves of Nicotiana tabacum L. by immunocytochemical methods. Plant Physiol, 1992, 100: 1591–1594
[9] Ishiyama K, Inoue E, Watanabe-Takahashi A, Obara M, Yamaya T, Takahashi H. Kinetic properties and ammonium-dependent regulation of cytosolic isoenzymes of glutamine synthetase in arabidopsis. J Biol Chem, 2004, 279: 16598–16605
[10]Llorca O, Betti M, González JM, Valencia A, Márquez AJ, Valpuesta JM. The three-dimensional structure of an eukaryotic glutamine synthetase: functional implications of its oligomeric structure. J Structural Biol, 2006, 156: 469–479
[11] Seabra AR, Vieira CP, Cullimore JV, Carvalho HG.Medicago truncatula contains a second gene encoding a plastid located glutamine synthetase exclusively expressed in developing seeds. BMC Plant Biol, 2010, 10: 183
[12] Singh K K, Ghosh S. Regulation of glutamine synthetase isoforms in two differentially drought-tolerant rice (Oryza sativa L.) cultivars under water deficit conditions. Plant Cell Rep, 2013, 32: 183–193
[13] Martin A, Lee J, Kichey T, Gerentes D, Zivy M, Tatout C, Dubois F, Balliau T, Valot B, Davanture M, Tercé-Laforgue T, Quilleré I, Coque M, Gallais A, Gonzalez-Moro MB, Bethencourt L, Habash DZ, Lea PJ, Charcosset A, Perez P, Murigneux A, Sakakibara H, Edwards KJ, Hirel B. Two cytosolic glutamine synthetase isoforms of maize are specifically involved in the control of grain production. Plant Cell, 2006, 18: 3252–3274
[14] McParland RH, Guevara JG, Becker RR, Evans HJ. The purification and properties of the glutamine synthetase from the cytosol of soya-bean root nodules. Biochem J, 1976, 153: 597–606
[15] Unno H, Uchida T, Sugawara H, Kurisu G, Sugiyama T, Yamaya T, Sakakibara H, Hase T, Kusunoki M. Atomic structure of plant glutamine synthetase: a key enzyme for plant productivity. J Biol Chem, 2006, 281: 29287–29296
[16] Seabra A R, Carvalho H, Pereira P J. Crystallization and preliminary crystallographic characterization of glutamine synthetase from Medicago truncatula. Acta Crystallographica, 2009, 65: 1309–1312
[17] Torreira E, Seabra AR, Marriott H, Zhou M, Llorca Ó, Robinson CV, Carvalho HG, Fernández-Tornero C, Pereira PJ. The structures of cytosolic and plastid-located glutamine synthetases from Medicago truncatula reveal a common and dynamic architecture. Acta Crystallographica, 2014, 70: 981–993
[18] Wittig I, Braun H P, Schägger H. Blue native PAGE. Nat Protocols, 2006, 1: 418–428
[19] Wittig I, Karas M, Schägger H. High resolution clear native electrophoresis for in-gel functional assays and fluorescence studies of membrane protein complexes. Molecular & Cellular Proteomics, 2007, 6: 1215–1225
[20] Kimata-Ariga Y, Hase T. Multiple complexes of nitrogen assimilatory enzymes in spinach chloroplasts: possible mechanisms for the regulation of enzyme function. PLoS One, 2014, 9(10): e108965
[21] Seabra A R, Silva L S, Carvalho H G. Novel aspects of glutamine synthetase (GS) regulation revealed by a detailed expression analysis of the entire GS gene family of Medicago truncatula under different physiological conditions. BMC Plant Biol, 2013, 13: 137–142
[22] Wang X C, Wei Y H, Shi L X, Ma X M, Theg SM. New isoforms and assembly of glutamine synthetase in the leaf of wheat (Triticum aestivum L.). J Exp Bot, 2015, 66: 6827–6834
[23] Mann A F, Fentem P A, Stewart G R. Identification of two forms of glutamine synthetase in barley (Hordeum vulgare). Biochem Biophys Res Commun, 1979, 88: 515–521
[24] 郭玉朋. 植物光呼吸途径研究进展. 草业学报, 2014, 23: 322–329
Guo Y P.A study on advances in plant photorespiration. Acta Pratac Sin, 2014, 23: 322–329 (in Chinese with English abstract)
[25] Bernard SM, Møller AL, Dionisio G, Kichey T, Jahn TP, Dubois F, Baudo M, Lopes MS, Tercé-Laforgue T, Foyer CH, Parry MA, Forde BG, Araus JL, Hirel B, Schjoerring JK, Habash DZ. Gene expression, cellular localisation and function of glutamine synthetase isozymes in wheat (Triticum aestivum L.). Plant Mol Biol, 2008, 67: 89–105
[26] Hirel B, Andrieu B, Valadier M H,Renarda S, Quilleré I,Chelleb M, Pommel B, Fournier C, Drouetb J L. Physiology of maize: II.identification of physiological markers representative of the nitrogen status of maize (Zea mays) leaves during grain filling. Physiol Plant, 2005, 124: 178–188

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